Oxidative stress has previously been shown to play a role in the pathogenesis of diabetes mellitus (DM) and its complications. In the present study, the effects of supplementation with dietary antioxidants, flaxseed and α-tocopherol were investigated in diabetic golden Syrian hamsters fed with a high-fat diet. Thirty-five golden Syrian hamsters were randomly divided into a control group (C) and four diabetic groups (DM, DM + flax, DM + E and DM + Flax + E). The hamsters received four different diets for a 20-week period, as follows: i) Groups C and DM received a high-fat diet (40% energy as fat), deficient in α-linolenic acid (ALA); ii) the DM + Flax group received a high-fat diet enriched with ground flaxseed 15 g/100 g of food, rich in ALA; iii) the DM + E group received a high-fat diet enriched with vitamin E, 40 mg α-tocopherol/100 g of food; and iv) the DM + Flax + E group received a high-fat diet enriched with flaxseed and vitamin E. The results of serum lipid and oxidative stress analysis suggested that the antiatherogenic effect of flaxseed, α-tocopherol and their combination added to a high-fat diet in diabetic hamsters was based primarily on their antioxidative role, demonstrated by decreased serum lipid peroxidation and increased liver glutathione content. Improvements of serum glucose and non-high-density lipoprotein cholesterol (HDL-C) levels were observed and may have contributed to the prevention of diabetic macroangiopathy evidenced in the histopathological examination. The antioxidant effect of flaxseed was similar to that of α-tocopherol in diabetic hamsters fed a high-fat diet and combined supplementation did not appear to bring more benefits than flaxseed alone. Moreover, the high dose of ground flaxseed alone may have a better cardioprotective effect than α-tocopherol in diabetic hamsters by reducing total cholesterol and non-HDL-C levels and increasing HDL-C levels.